Santin A D, Hermonat P L, Ravaggi A, Bellone S, Pecorelli S, Cannon M J, Parham G P
Division of Gynecologic Oncology, Department of Obstetrics and Gynecology, University of Arkansas, Little Rock, AR 72205-7199, USA.
Am J Obstet Gynecol. 2000 Sep;183(3):601-9. doi: 10.1067/mob.2000.107097.
The purpose of this study was to evaluate the potential of dendritic cells pulsed with whole-tumor extracts derived from autologous ovarian cancer cells in eliciting a tumor-specific cytotoxic T-cell response in vitro from patients with advanced ovarian cancer.
CD8(+) T lymphocytes stimulated in vitro with autologous ovarian tumor lysate-pulsed dendritic cells were tested for their ability to induce a human leukocyte antigen class I-restricted cytotoxic T-lymphocyte response able to specifically kill autologous tumor cells in standard 6-hour chromium 51 cytotoxicity assays. In addition, to correlate cytotoxic activity by cytotoxic T-lymphocytes with a particular lymphoid subset, 2-color flow cytometric analysis of intracellular cytokine expression (interferon gamma and interleukin 4) at the single-cell level was performed.
Cytotoxic T lymphocytes specific for autologous ovarian tumor cells were elicited from 3 patients with advanced ovarian cancer. Although cytotoxic T-lymphocyte populations expressed strong cytolytic activity against autologous tumor cells, they did not lyse concanavalin A-stimulated autologous lymphocytes or autologous Epstein-Barr virus-transformed lymphoblastoid cell lines and showed negligible cytotoxicity against the natural killer cell-sensitive cell line K-562. Cytotoxic effect against the autologous tumor cells was inhibited by an anti-human leukocyte antigen class I monoclonal antibody (W6/32). It is interesting that CD8(+) cytotoxic T lymphocytes expressed variable levels of CD56, a marker that may be associated with high cytotoxic activity. Finally, most of the tumor-specific CD8(+) T cells exhibited a T(H)1 cytokine bias, and a high percentage of interferon gamma expressors among cytotoxic T lymphocytes was correlated with higher cytotoxic activity.
These data show that tumor lysate-pulsed dendritic cells can consistently induce in vitro specific CD8(+) cytotoxic T lymphocytes able to kill autologous tumor cells from patients with advanced stage ovarian cancer. This novel approach may have important implications for the treatment of residual or resistant disease with active or adoptive immunotherapy after standard surgical and cytotoxic treatment.
本研究旨在评估用源自自体卵巢癌细胞的全肿瘤提取物脉冲处理的树突状细胞在体外诱导晚期卵巢癌患者产生肿瘤特异性细胞毒性T细胞反应的潜力。
在标准的6小时铬51细胞毒性试验中，检测用自体卵巢肿瘤裂解物脉冲处理的树突状细胞体外刺激的CD8(+) T淋巴细胞诱导人类白细胞抗原I类限制性细胞毒性T淋巴细胞反应以特异性杀伤自体肿瘤细胞的能力。此外，为了将细胞毒性T淋巴细胞的细胞毒性活性与特定淋巴细胞亚群相关联，在单细胞水平上对细胞内细胞因子表达（干扰素γ和白细胞介素4）进行双色流式细胞术分析。
从3例晚期卵巢癌患者中诱导出了对自体卵巢肿瘤细胞具有特异性的细胞毒性T淋巴细胞。尽管细胞毒性T淋巴细胞群体对自体肿瘤细胞表现出强大的溶细胞活性，但它们不裂解伴刀豆球蛋白A刺激的自体淋巴细胞或自体爱泼斯坦-巴尔病毒转化的淋巴母细胞系，并且对自然杀伤细胞敏感细胞系K-562的细胞毒性可忽略不计。抗人类白细胞抗原I类单克隆抗体（W6/32）可抑制对自体肿瘤细胞的细胞毒性作用。有趣的是，CD8(+) 细胞毒性T淋巴细胞表达可变水平的CD56，这是一种可能与高细胞毒性活性相关的标志物。最后，大多数肿瘤特异性CD8(+) T细胞表现出TH1细胞因子偏向，并且细胞毒性T淋巴细胞中高百分比的干扰素γ表达者与更高的细胞毒性活性相关。
这些数据表明，肿瘤裂解物脉冲处理的树突状细胞可在体外持续诱导能够杀伤晚期卵巢癌患者自体肿瘤细胞的特异性CD8(+) 细胞毒性T淋巴细胞。这种新方法可能对标准手术和细胞毒性治疗后采用主动或过继性免疫疗法治疗残留或耐药疾病具有重要意义。
